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In Vitro Antibacterial Effects of the Chimeric Peptides from Chicken and Pig Antimicrobial Peptide NK-Lysin (닭과 돼지의 항균펩타이드 NK-Lysin으로부터 조합된 펩타이드의 In Vitro 항균효과)

  • Hong, Yeojin;Lee, Gi Yong;Yang, Soo-Jin;Lillehoj, Hyun Soon;Hong, Yeong Ho
    • Korean Journal of Poultry Science
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    • v.49 no.2
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    • pp.69-77
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    • 2022
  • Antimicrobial peptides (AMPs) play an important role in innate immunity against pathogenic infections. AMPs exterminate pathogenic bacteria by disrupting cell membranes or inhibiting intracellular molecules. NK-2, first identified in pigs and derived from NK-lysin, has antimicrobial effects against bacteria and parasites. In this study, chimeric peptides (cpNK) of chicken and pig NK-2 and cpNK-derived peptides (cpNK-a1 and cpNK-a2) were synthesized, and their antimicrobial effects against various pathogenic bacteria such as Escherichia coli, Salmonella spp., Listeria monocytogenes, Staphylococcus aureus, and methicillin-resistant Staphylococcus aureus (MRSA) were investigated. The structure of chimeric peptides from chicken and pig NK-2, cpNK, include α-helix like NK-2 and peptide net charge was +9 like porcine NK-2. The cpNK peptide showed powerful bactericidal effects against most bacterial species, including MRSA, especially against gram-negative bacteria. Furthermore, cpNK-derived short peptides, cpNK-a1 and a2 also showed bactericidal activity, but the effects were weaker than those of cpNK. Therefore, we conclude that cpNK- and cpNK-derived short peptides have the potential to be used as antibiotic alternatives.

SCIATIC NERVE REGENERATION USING CALCIUM PHOSPHATE COATED CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE-TRANSFECTED SCHWANN CELL IN RAT (인회석 박막 피복 도관과 Brain-derived neurotrophic factor(BDNF) 유전자 이입 슈반세포를 이용한 백서 좌골신경 재생에 관한 연구)

  • Choi, Won-Jae;Ahn, Kang-Min;Hwang, Soon-Jeong;Choung, Pill-Hoon;Kim, Myung-Jin;Kim, Nam-Yeol;Yoo, Sang-Bae;Jahng, Jeong-Won;Kim, Hyun-Man;Kim, Joong-Soo;Kim, Yun-Hee;Kim, Soung-Min;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.3
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    • pp.199-218
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    • 2005
  • Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.

Comparison of Disk Tension Infiltrometer and van Genuchten-Mualem Model on Estimation of Unsaturated Hydraulic Conductivity (장력 침투계(Disk Tension Infiltrometer)와 van Genuchten-Mualem 모형 적용에 따른 불포화수리 전도도의 비교 해석)

  • Hur, Seung-Oh;Jung, Kang-Ho;Park, Chan-Won;Ha, Sang-Keun;Kim, Geong-Gyu
    • Korean Journal of Soil Science and Fertilizer
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    • v.39 no.5
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    • pp.259-267
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    • 2006
  • Hydraulic conductivity is the rate of water flux on hydraulic gradient. The van Genuchten Mualem (VGM) model is frequently used for describing unsaturated state of soils, that is composed with the function of soil water potential and soil water content and requests various parameters. This study is to get the value of VGM parameters used Rosetta computer program based on neural network analysis method and to calculate VGM parameters. VGM parameters included Ko(effective saturated hydraulic conductivity), ${\theta}r$(residual soil water content), ${\theta}s$(saturated soil water content), L, n and m. The unsaturated hydraulic conductivity at 10 kPa was calculated by using Rosetta program. Unsaturated hydraulic conductivities of 17 soil series at 1, 3, 5, 7 kPa were also obtained by applying saturated hydraulic conductivity by disk tension infiltrometer based on Gardner and Wooding's equation. Water flow at the water potential of 3 kPa was very low except Namgye, Hagog, Baegsan, Sangju, Seogcheon, Yesan soil series. Unsaturated hydraulic conductivity at 1 kPa showed the highest value for Samgag soil series and was in order of Yesan, Hwabong, Hagog and Baegsan soil series. Those of Gacheon, Seocheon and Ugog soil series were very low. When the value by VGM was compared with the value by disc tension infiltrometer, there was a tendency with exponential function to soils without gravel but there was no tendency to soils including gravel. Conclusively, it would be limited that VGM model for unsaturated hydraulic conductivity analysis applies to Korean agricultural land including gravel and having steep slope, shallow soil depth.